Hematopoietic neoplasm chemotherapy

ABSTRACT

A method and medicament for treating mixed lineage leukemia; translocated mixed lineage leukemia; translocated mixed lineage leukemia based acute myelogenous leukemia; translocated mixed lineage leukemia based acute lymphoid leukemia; a non-MLL based chronic myeloproliferative disorder, or non-MLL based acute lymphoid leukemia is provided.

Leukemia's, myeloid, lymphoid and mixed lineage, are clonal neoplasmsthat arise as a result of at least one chromosomal abnormality. Theseabnormalities result in a change in gene structure and function.Treatment regimens generally comprise several chemotherapeutic agentsadministered concomitantly or sequentially. Recent advances, such asimatinib mesylate, nilotinib and dasatinib, have improved the time toprogression and overall survival in chronic myeloid leukemia patients.Despite these advances, the therapeutic effectiveness of a particularagent, or combination of agents, is frequently not sustained asadditional genetic and/or epigenetic abnormalities are acquired. Moreefficacious chemotherapeutic agents for the treatment of chronic myeloidleukemia and other hematopoietic malignancies are desirable.

Glycogen synthase kinase 3 (GSK3) is a serine/threonine kinaseconstitutively active in normal resting cells and is regulated throughinhibition of its activity. GSK3 is implicated in various signaltransduction networks known to regulate a variety of cell functions.Abnormalities in pathways that use GSK3 as a regulator are implicated indisease pathogenesis which has prompted efforts to develop GSK3 specificinhibitors for various therapeutic applications such asnon-insulin-dependent-diabetes, Alzheimer's disease and otherneurodegenerative disorders, and developmental disorders. Due to itsinvolvement in multiple pathways, suitable potency of GSK3 inhibition isan important factor in the development of inhibitors for therapeuticapplications.

Recently, a specific GSK3 inhibitor has been reported to potentiate theeffects of specified chemotherapeutic agents at particular solid tumortypes, although lacking useful antitumor activity in its own right,WO2009/006043.

It has also been disclosed that GSK3 plays a role in the maintenance ofgenetically defined translocated mixed lineage leukemia (MLL leukemia).Wang et al., Nature, 455, 1205-1210 (2008). This same report alsodiscloses GSK3 inhibition in genetically defined translocated MLLleukemia by specific GSK3 inhibitor compounds. The GSK-3 inhibitor[3-(2,4-dichlorophenyl)-4-(1-methyl-1H-indol-3-yl)-1H-pyrrole-2,5-dione(SB216763), and GSK-3 Inhibitor IX, (2′Z,3′E)-6-bromoindirubin-3′-oxime(“GSK3-IX”) are mentioned as evidencing positive results.

There is a need for leukemia selective chemotherapeutic agents thatexhibit per se therapeutic activity, and improved efficacy in thetreatment of a leukemia patient with a specific type of leukemia. TheGSK3β inhibitor7-(2,5-dihydro-4-imidazo[1,2-a]-pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinyl-carbonyl)pyrrolo[3,2,1-jk][1,4]benzodiazepineevidences selectivity, per se therapeutic activity, and improvedefficacy over SB216763 and GSK3-IX, against several types of leukemia.

One aspect of the invention provides a method of treating a patientsuffering from mixed lineage leukemia; translocated mixed lineageleukemia; translocated mixed lineage leukemia based acute myelogenousleukemia; translocated mixed lineage leukemia based acute lymphoidleukemia; a non-MLL based chronic myeloproliferative disorder; or anon-MLL based acute lymphoid leukemia comprising administering to aleukemia patient in need of such treatment an effective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof.

A second aspect of the invention provides a method of treating a patientsuffering from mixed lineage leukemia; translocated mixed lineageleukemia; translocated mixed lineage leukemia based acute myelogenousleukemia; or translocated mixed lineage leukemia based acute lymphoidleukemia comprising administering to a leukemia patient in need of suchtreatment an effective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof.

A third aspect of the invention provides a method of treating a patientsuffering from mixed lineage leukemia comprising administering to aleukemia patient in need of such treatment an effective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof.

A fourth aspect of the invention provides a method of treating a patientsuffering from translocated mixed lineage leukemia comprisingadministering to a leukemia patient in need of such treatment aneffective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof.

A fifth aspect of the invention provides a method of treating a patientsuffering from translocated mixed lineage leukemia based acutemyelogenous leukemia comprising administering to a leukemia patient inneed of such treatment an effective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof.

A sixth aspect of the invention provides a method of treating a patientsuffering from translocated mixed lineage leukemia based acute lymphoidleukemia comprising administering to a leukemia patient in need of suchtreatment an effective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof.

A seventh aspect of the invention provides a method of treating apatient suffering from a non-MLL based chronic myeloproliferativedisorder comprising administering to a leukemia patient in need of suchtreatment an effective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof.

An eighth aspect of the invention provides a method of treating apatient suffering from non-MLL based acute myelogenous leukemia;erythroleukemia; or chronic myelogenous leukemia comprisingadministering to a leukemia patient in need of such treatment aneffective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof.

A ninth aspect of the invention provides a method of treating a patientsuffering from non-MLL based erythroleukemia comprising administering toa leukemia patient in need of such treatment an effective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof.

A tenth aspect of the invention provides a method of treating a patientsuffering from non-MLL based chronic myelogenous leukemia comprisingadministering to a leukemia patient in need of such treatment aneffective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof.

An eleventh aspect of the invention provides a method of treating apatient suffering from non-MLL based acute myelogenous leukemiacomprising administering to a leukemia patient in need of such treatmentan effective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof.

A twelfth aspect of the invention provides a method of treating apatient suffering from non-MLL based acute lymphoid leukemia comprisingadministering to a patient in need of such treatment an effective amountof7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof.

A thirteenth aspect of the invention provides a method of treating apatient suffering from a non-MLL based JAK2 (+) chronicmyeloproliferative disorder comprising administering to a leukemiapatient in need of such treatment an effective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof.

A fourteenth aspect of the invention provides a method of treating apatient suffering from non-MLL based Philadelphia positive chronicmyelogenous leukemia comprising administering to a patient in need ofsuch treatment an effective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof.

A fifteenth aspect of the invention provides the use of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof for thepreparation of a medicament for the treatment of mixed lineage leukemia;translocated mixed lineage leukemia; translocated mixed lineage leukemiabased acute myelogenous leukemia; translocated mixed lineage leukemiabased acute lymphoid leukemia; a non-MLL based chronicmyeloproliferative disorder or a non-MLL based acute lymphoid leukemia.

A sixteenth aspect of the invention provides the use of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof for thepreparation of a medicament for the treatment of mixed lineage leukemia;translocated mixed lineage leukemia; translocated mixed lineage leukemiabased acute myelogenous leukemia; or translocated mixed lineage leukemiabased acute lymphoid leukemia.

A seventeenth aspect of the invention provides the use of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof for thepreparation of a medicament for the treatment of mixed lineage leukemia.

An eighteenth aspect of the invention provides the use of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof for thepreparation of a medicament for the treatment of translocated mixedlineage leukemia.

A nineteenth aspect of the invention provides the use of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof for thepreparation of a medicament for the treatment of translocated mixedlineage leukemia based acute myelogenous leukemia.

A twentieth aspect of the invention provides the use of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof for thepreparation of a medicament for the treatment of translocated mixedlineage leukemia based acute lymphoid leukemia.

A twenty-first aspect of the invention provides the use of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof for thepreparation of a medicament for the treatment of a non-MLL based chronicmyeloproliferative disorder.

A twenty-second aspect of the invention provides the use of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof for thepreparation of a medicament for the treatment of non-MLL based acutemyelogenous leukemia; erythroleukemia; or chronic myelogenous leukemia

A twenty-third aspect of the invention provides the use of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof for thepreparation of a medicament for the treatment of non-MLL basederythroleukemia.

A twenty-fourth aspect of the invention provides the use of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof for thepreparation of a medicament for the treatment of non-MLL based chronicmyelogenous leukemia.

A twenty-fifth aspect of the invention provides the use of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof for thepreparation of a medicament for the treatment of non-MLL based acutemyelogenous leukemia.

A twenty-sixth aspect of the invention provides the use of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof for thepreparation of a medicament for the treatment of a non-MLL based JAK2(+) chronic myeloproliferative disorder.

A twenty-seventh aspect of the invention provides the use of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof for thepreparation of a medicament for the treatment of non-MLL basedPhiladelphia positive chronic myelogenous leukemia.

A twenty-eighth aspect of the invention provides the use of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof for thepreparation of a medicament for the treatment of non-MLL based acutelymphoid leukemia.

A twenty-ninth aspect of the invention provides a compound7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or solvate thereof, for use in thetreatment of mixed lineage leukemia; translocated mixed lineageleukemia; translocated mixed lineage leukemia based acute myelogenousleukemia; translocated mixed lineage leukemia based acute lymphoidleukemia; a non-MLL based chronic myeloproliferative disorder; or anon-MLL based acute lymphoid leukemia. In a particular embodiment, theleukemia is translocated mixed lineage leukemia based acute myelogenousleukemia; translocated mixed lineage leukemia based acute lymphoidleukemia; non-MLL based chronic myeloproliferative disorder selectedfrom non-MLL based acute myelogenous leukemia, erythroleukemia, orchronic myelogenous leukemia; non-MLL based acute myelogenous leukemia;or non-MLL based chronic myelogenous leukemia.

The compound7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinyl-carbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineis taught to be an inhibitor of GSK-3β in WO 03/076442, where it isreferred to as3-(9-fluoro-6-(piperidin-1-yl)carbonyl)-6,7-dihydro-6H-[1,4]diazepino-[6,7,1-hi]indol-1-yl)-4-(imidazo[1,2-a]pyridin-3-yl)-2,5-dioxopyrrole(Example 365, page 113). The two naming conventions described above aretaken to be synonymous and each is taken to identify the followingstructure:

Compound 1 is a base, and accordingly may react with any of a number ofinorganic and organic acids to form pharmaceutically acceptable acidaddition salts. Pharmaceutically acceptable acid addition salts of thecompound of the present invention and common methodology for preparingthem are well known in the art. See, e.g., P. Stahl, et al., Handbook ofPharmaceutical Salts: Properties, Selection and Use, (VCHA/Wiley-VCH,2002); S. M. Berge, et al., “Pharmaceutical Salts,” Journal ofPharmaceutical Sciences, Vol. 66, No. 1, January 1977. Preferredpharmaceutically acceptable acids include HCl, HBr, sulfuric acid andmethanesulfonic acid.

Compound 1 forms solvates with, for example, water (hydrate anddihydrate), methanol, and ethanol. A preferred solvate is that formedwith ethanol.

As used herein, the term “patient” means mammal; “mammal” means theMammalia class of higher vertebrates; and the term “mammal” includes,but is not limited to, a human. The preferred patient is a human.

As used herein, the terms “myeloid” and “myelogenous” are usedinterchangeably. Similarly, “lymphoid” and “lymphogenous” are usedinterchangeably.

Also as used herein, the term “per se” means independent therapeuticpotency. There is no requirement for coadministration of a second activeoncologic chemotherapeutic agent to obtain or potentiate leukemiatreatment efficacy although such coadministration may be desirable.

There is considerable variability in the degree to which cancer genomesare aberrant at the chromosomal level. Some cancers are characterized bya single signature chromosomal aberration while others have numerousaberrations and very complex karyotypes. In solid tumors, such asepithelial-derived, cytogenetic analyses have identified many structuralchromosomal aberrations. This is in contrast to hematopoieticmalignancies where a relative few are causally linked and recurrent. Themajority of recurrent chromosomal aberrations are found in hematopoieticmalignancies in contrast to solid tumors. Deletion and amplification aremore characteristic of solid tumors, along with progressive geneticinstability and the acquisition of a complex panoply of genomicaberrations in contrast to hematopoietic malignancies.

“Non-MLL based chronic myeloproliferative disorders” are acquired clonalabnormalities of the hematopoietic stem cell and include polycythemiavera, myelofibrosis, essential thrombocytosis, chronic myeloid leukemia,myelodysplastic syndrome and acute myeloid leukemia and includeserythroleukemia. Since the stem cell gives rise to myeloid, erythroid,and platelet cells, qualitative and quantitative changes may be seen inone, two or all those cell lines depending on where in the maturationprocess from the pluripotent stem cell to a dedicated cell typeprogenitor stem cell the abnormality occurs. In some disorders (such aschronic myeloid leukemia), specific characteristic chromosomal changesare seen. Chronic myeloproliferative disorders produce characteristicsyndromes with defined clinical and laboratory features.

Non-MLL based polycythemia vera causes overproduction of all threehematopoietic cell lines, most prominently erythroid cells. Erythroidproduction is independent of erythropoietin. A mutation in Janus kinase2, chromosome band 9p24, (JAK2 (+)), a cell signaling molecule isbelieved involved in the pathogenesis and is a criteria of diagnosis.

Non-MLL based myelofibrosis is characterized by fibrosis of the bonemarrow, splenomegaly, and a leukoerythroblastic peripheral blood picturewith teardrop poikilocytosis. In response to bone marrow fibrosis,extramedullary hematopoiesis takes place in the liver, spleen, and lymphnodes. Abnormalities of JAK2 (JAK2 (+)) and its signaling pathway arebelieved involved in the pathogenesis.

Non-MLL based essential thrombocytosis is characterized by markedproliferation of the megakaryocytes in the bone marrow leading to anelevated platelet count. A high frequency of JAK2 mutations (JAK2 (+))has been seen in patients and is believed involved in the pathogenesis.

Non-MLL based chronic myeloid leukemia (CML) is characterized byoverproduction of myeloid cells. These myeloid cells retain the capacityfor differentiation and normal bone marrow function is retained duringthe early phases. CML is frequently characterized by a specificchromosomal abnormality and specific molecular abnormality. ThePhiladelphia chromosome is a reciprocal translocation between the longarms of chromosomes 9 and 22. A large portion of 22q is translocated to9q, and a smaller piece of 9q is moved to 22q. The portion of 9q that istranslocated contains abl, a protooncogene that is the cellular homologof the Ableson murine leukemia virus. The abl gene is received at aspecific site on 22q, the break point cluster (bcr). The fusion genebcr/abl produces a novel protein that differs from the normal transcriptof the abl gene in that it possesses tyrosine kinase activity. Evidencethat the bcr/abl fusion gene is pathogenic is provided by transgenicmouse models in which introduction of the gene almost invariably leadsto leukemia. The presence of this translocation is referred to asPhiladelphia positive. In early CML (chronic phase) normal bone marrowfunction is retained, white blood cells differentiate and, despite somequalitative abnormalities, the neutrophils combat infection normally.CML, however, is inherently unstable and without treatment progresses toan accelerated phase and then an acute or blast phase which ismorphologically indistinguishable from conventional acute myeloidleukemia. This progression has been associated with the acquisition ofadditional genetic and/or epigenetic abnormalities.

Non-MLL based myelodysplastic syndromes are a group of acquired clonaldisorders of the hematopoietic stem cell. They are characterized bycytopenia, a hypercellular marrow, and a number of morphologic andcytologic abnormalities. Typically, morphologic abnormalities arepresent in two or more hematopoietic cell lines. These disorders aretypically idiopathic but may be seen after cytotoxic chemotherapy.Although no single specific chromosomal abnormality is seen inmyelodysplasia, there are frequent abnormalities involving the long armof chromosome 5 as well as deletions of chromosomes 5 and 7. Non-MLLbased myelodysplasia with a proliferative syndrome are termed chronicmyelomonocytic leukemia (CMML).

Non-MLL based acute myeloid leukemia (AML) is a malignancy of one ormore myeloid hematopoietic progenitor cells not based upon MLLleukemogenesis. These cells proliferate in an uncontrolled fashion andreplace normal bone marrow elements. Although most cases arise with noclear cause, radiation and some toxins are leukemogenic. In addition, anumber of chemotherapeutic agents may cause leukemia. The leukemia'sseen after toxin or chemotherapy exposure are often associated withabnormalities in chromosomes 5 and 7 or chromosome 11q23. The mostcommon cytogenetic abnormalities causally linked to non-MLL based AMLare t(8;21)(q22;q22) affording the AML1/ETO fusion gene; Inv(16)(p13q22)affording the CBFβ/MYH11 fusion gene; t(16;16)(p13;q22),t(15;17)(q21;q11), t(11;17)(q23;q11), t(5;17)(q35;q12-21),t(11;17)(q13;q21), and t(17;17)(q11;q21) affording various RARαcontaining fusion genes; 5/5q-; -7/7q-; 17p abn or i(17q); del(20q);dmins hsrs; +13; Inv(3)(q21q26), and t(3;3)(q21;q26) affording theRibophorin/EVI1 fusion gene. The Auer rod, an eosinophilic needle-likeinclusion in the cytoplasm, is pathognomonic of non-MLL based acutemyeloid leukemia (AML). Leukemia cells retain properties of the lineagesfrom which they are derived or based. AML cells usually express myeloidantigens such as CD13 or CD33.

Non-MLL based acute lymphoid leukemia (ALL) is a malignancy of thelymphoid hematopoietic progenitor cell not based upon MLLleukemogenesis. As noted above, leukemia cells retain properties of thelineages from which they are derived or based. Non-MLL based ALL cellsof B lineage will express lymphoid antigens such as CD19, common to allB cells, and most cases will express CD10 also known as common ALLantigen. Non-MLL based ALL cells of T lineage will usually not expressmature T-cell markers, such as CD3, 4, or 8, but will express somecombination of CD2, 5, and 7 and do not express surface immunoglobulin.Non-MLL based ALL cells frequently express terminal deoxynucleotidyltransferase (TdT). The most frequent recurrent genetic subtypes includeTEL-AML1; BCR-ABL; E2A/PBX1; IgH/MYC; numerous translocations involvingthe TCR ab (7q35) or TCR gd (14q11) loci; 1q deletions; SIL-SCL andNOTCH mutations.

Non-MLL based AML has been characterized in several ways. The FAB(French, American, British) classification is based on marrow morphologyand histochemistry as follows: acute undifferentiated leukemia (M0),acute myeloblastic leukemia (M1), acute myeloblastic leukemia withdifferentiation (M2), acute promyelocytic leukemia (APL) (M3), acutemyelomonocytic leukemia (M4), acute monoblastic leukemia (M5),erythroleukemia (M6), and megakaryoblastic leukemia (M7). The WorldHealth Organization has sponsored a classification of the leukemia's andother hematologic malignancies that incorporates cytogenetic, molecular,and immunophenotype information, International Classification ofDiseases for Oncology, Third edition, Percy et al., 2000.

Non-MLL based ALL may be classified by immunologic phenotype as follows:common, B cell, and T cell. As with non-MLL based AML, certain toxins,radiation and chemotherapeutic agents can cause non-MLL based ALL.

Mixed lineage leukemia (myeloid lymphoid leukemia; MLL) hascharacteristics of both non-MLL based AML and non-MLL based ALL. MLLspecifies a distinct gene expression profile over non-MLL based ALL andnon-MLL based AML; Armstrong et. al., Nature Genetics, 30, 41-47 (2002).MLL may result from recurrent chromosomal aberrations at chromosome 11at band q23 (MLL gene), chromosome fusions involving the long arm (q) ofchromosome 11 at band q23 with a gene from a different chromosomalregion, which may be translocated, or 11q23 may be internallyduplicated. Leukemia expressing MLL fusions are frequently aggressiveand resistant to chemotherapy. These fusions may translocate resultingin the MLL gene being rearranged. The MLL translocated gene fusions maycause either translocated MLL based AML or translocated MLL based ALL.For example, MLL-AF9 translocated gene fusions frequently, but notexclusively, cause AML (translocated MLL based AML). Other MLLtranslocated gene fusions associated with translocated MLL based AMLinclude MLL-AF10 and MLL-ELL. A translocated MLL gene fusion associatedwith translocated MLL based ALL is MLL-AF4.

Extensive catalogues of the cytogenetic aberrations in human cancer havebeen compiled and are maintained and regularly updated online (see TheMitelman Database of Chromosome Aberrations in Cancer at the US NationalCancer Institute (NCI) Cancer Genome Anatomy Project (CGAP) Web site:http://cgap.nci.nih.gov). The database includes chromosomal aberrationsfor the hematopoietic malignancies of the present invention. TheWellcome Trust Sanger Institute Cancer Genome Project maintains adetailed online “Cancer Gene Census” of all human genes that have beencausally linked to tumorigenesis (seehttp://www.sanger.ac.uk/genetics/CGP/Census) as well as the COSMIC(Catalogue of Somatic Mutations in Cancer) database of somatic mutationsin human cancer (see http://www.sanger.ac.uk/genetics/CGP/cosmic). Afurther source containing abundant information on cytogenetic changescausally linked to leukemia's is the Atlas of Genetics and Cytogeneticsin Oncology and Haematology(http://atlasgeneticsoncology.org//Anomalies/Anomliste html#MDS). Thesedatabases also include chromosomal aberrations for the hematopoieticmalignancies of the present invention. An alternative source of theCancer Gene Census database is Holland-Frei Cancer Medicine, 7^(th) Ed.,(2006), Table 8-1 (See also Table 8-4 for the Most Frequent RecurrentChromosomal Abnormalities in Myeloid Disorders and Table 8-5 for theMost Frequent Recurrent Genetic Subtypes of B and T Cell ALL) and theCOSMIC database is Forbes et al., Br. J. Cancer, 2006, 94(2), 318-22.

Diagnosis of hematopoietic malignancies by complete blood counts, bonemarrow aspiration and biopsy, immunophenotyping and other tests areknown and routinely used. In addition to high resolution chromosomebanding and advanced chromosomal imaging technologies, chromosomeaberrations in suspected cases of hematopoietic malignancies can bedetermined through cytogenetic analysis such as fluorescence in situhybridization (FISH), karyotyping, spectral karyotyping (SKY), multiplexFISH (M-FISH), comparative genomic hybridization (CGH), singlenucleotide polymorphism arrays (SNP Chips) and other diagnostic andanalysis tests known and used by those skilled in the art.

Beyond the genetic chromosomal aberrations mentioned above, each of theleukemia's may also include epigenetic modifications of the genomeincluding DNA methylation, genomic imprinting, and histone modificationby acetylation, methylation, or phosphorylation. An epigeneticmodification may play an important role in the malignancy.

The phrase “an effective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor a pharmaceutically acceptable salt or a solvate thereof” is taken tomean the dosage of Compound 1 or a pharmaceutically acceptable salt or asolvate thereof necessary to either destroy the target leukemia cells orslow or arrest the progression of the leukemia in a patient. Anticipateddosages of Compound 1 or a pharmaceutically acceptable salt or a solvatethereof are in the range of 5 to 600 mg/patient/day. Preferred dosagesare anticipated to be in the range of 50 to 400 mg/patient/day. Mostpreferred dosages are anticipated to be in the range of 100 to 400mg/patient/day. The exact dosage required to treat a patient will bedetermined by a physician in view of the stage and severity of thedisease as well as the specific needs and response of the individualpatient.

The following in vitro and in vivo studies demonstrate the per setherapeutic activity and improved efficacy of Compound 1 against variousspecific leukemia cell lines.

In Vitro Efficacy Examples

Apoptosis or programmed cell death is characterized by a set ofbiochemical reactions, one of which is the induction of caspases.Activated caspases are proteases that participate in a cascade ofcleavage events that disable key enzymes responsible for cellhomeostasis and repair. Caspases 3 and 7 play key effector roles inapoptosis and can be detected and measured by a fluorescent biochemicalassay. The increase of Caspase-3/7 activity in cells is directlycorrelated to apoptotic activity. (D. W. Nicholson, et al., Nature, 376,37-43 (1995)) The Promega Apo-ONE Homogeneous Caspase-3/7 Assay Kit isused (Catalog #G7791). The assay buffer consists of 30 mM HEPES(N-(2-hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid) pH 7.4, 150 mMNaCl, 50 mM KCl, 10 mM MgCl₂, 0.4 mM EGTA (ethylene glycol tetraaceticacid), 0.5% Nonidet P40 (octylphenolpoly(ethyleneglycol ether)), 0.1%CHAPS (3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate hydrateand 10% sucrose, which lyses/permeabilizes cultured cells and a caspase3/7 substrate, Z-DEVD (Z-Asp(OMe)-Glu(OMe)-Val-Asp(OMe)), coupled to aprofluorescent rhodamine 110. When the buffer-substrate mixture is addedto a test sample, the cleavage and subsequent removal of the DEVDpeptides by caspase 3/7 activity results in intense fluorescence of therhodamine 110 leaving group, which is detected by excitation at 490 nm.The amount of fluorescent product is proportional to the amount ofcaspase 3/7 cleavage activity in the sample.

To measure the apoptotic effect of test compounds, tumor cells areplated at 1×10⁴ cells per well in 96 well plates and incubated overnightat 37° C., with 5% CO₂. Tumor cells are treated with test compound atdesired concentrations in triplicate, including untreated/negativecontrol wells. The assay plates are re-incubated for 48 hrs. At the endof the incubation period, a mixture of the assay buffer and substrate isadded to each sample well. The fluorescence in each well is measured atan excitation wavelength of 480+/−20 nm and an emission wavelength of530+/−25 nm. The % increase of caspase activity in treated cells iscalculated relative to untreated controls.

Cell viability is determined by the CellTiter-Glo® Luminescent CellViability Assay (Promega, Catalog #G7570) which is a method ofestimating viable cell number based on quantitation of ATP inmetabolically active cells. After cells are lysed, the mono-oxygenationof the substrate luciferin is catalyzed by the enzyme luciferase in thepresence of Mg2+, ATP and molecular oxygen, resulting in the generationof a luminescent signal that is proportional to the number of viablecells in the assay wells.

To measure the viability of cells after treatment with compounds, tumorcells are plated at 2×10⁴ per well in 96 well plates and incubatedovernight at 37° C., with 5% CO2. Tumor cells are treated with testcompound at desired concentrations in triplicate, includinguntreated/negative control cells. The assay plates are re-incubated for48 hrs. At the end of the incubation period, a mixture of lysis assaybuffer and substrate is added to each sample well. The luminescence ineach well is measured using a microtiter plate luminometer.

MV4;11 is a human acute myeloid leukemia line characterized by thepresence of a fusion transcript comprised of the MLL and AF4 genes andby the presence of an internal tandem duplication in the juxtamembraneregion of the FLT-3 gene. RS4;11 is a human acute lymphoid leukemia cellline characterized by the presence of a fusion transcript comprised ofthe MLL and AF4 genes. REH is a human acute lymphoid leukemia (non-T;non-B) cell line characterized by the presence of a fusion transcriptcomprised of the TEL and AML1 genes. Kasumi 1 is a human acute myeloidleukemia line characterized by the presence of a fusion transcriptcomprised of the AML1 and ETO genes. K562 is a human chronic myelogenousleukemia cell line characterized by the presence of a fusion transcriptcomprised of the Bcr and Abl genes. HEL 92.1.7 is a humanerythroleukemia cell line characterized by the presence of a V617Fmutation in the JAK2 gene. Jurkatt is a human acute T-cell leukemia cellline. Each of the cell lines are obtained from the American Type CultureCollection (ATCC). In the following tables the term “Compound 1” or“Cmpd 1” means7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinyl-carbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepine.The GSK-3 inhibitor[(3-(2,4-dichlorophenyl)-4-(1-methyl-1H-indol-3-yl)-1H-pyrrole-2,5-dione(SB216763), Sigma-Aldrich is used as a positive comparator control insome experiments. The compound GSK-3 Inhibitor IX,(2′Z,3′E)-6-bromoindirubin-3′-oxime (“GSK3-IX”) Calbiochem, is used as apositive control in some experiments. Both SB216763 and GSK3-IX arementioned in the Wang et al., Nature, 455, 1205-1210 (2008) paper asevidencing positive results.

The data in Table 1 are expressed as % increase of caspase 3 activityrelative to untreated controls unless otherwise noted.

TABLE 1 Caspase 3 Activity % % Increase Increase in in % Caspase 3Caspase 3 Increase after after in Concen- treatment Concen- treatmentCaspase 3 tration with tration with activity of Cmpd 1 of SB216763 withCompound (mean of SB216763 (mean of Cell line vehicle 1 (in μM)triplicates (in μM) triplicates MV4; 11 0 0.010 188 30 83 RS4; 11 00.009 111 20 117 REH 0 0.0007 132 20 113 Kasumi 1 0 0.370 133 10 27 HEL0 0.120 284 10 250 92.1.7 K562 0 0.0007 1057 20 223 Jurkat 0 0.370 14010 101

The data in Table 1 evidences per se activity by Compound 1 against allcell lines tested and particularly against non-MLL based AML, CML,erythroleukemia and ALL. The data also evidences improved efficacy ofCompound 1 over SB216763 against all cell lines tested.

The data in Table 2 are expressed as the estimated concentrationrequired for a reduction in cell viability by 50% (EC50) after treatmentwith Compound 1 or SB216763.

TABLE 2 Reduction in Cell Viability Reduction in Reduction in cellReduction in cell cell viability viability after viability after aftertreatment with treatment with treatment with Cmpd 1 SB216763 - EC50GSK3-IX - EC50 EC50 in μM in μM (mean of in μM (mean of Cell line (meanof triplicates) triplicates) triplicates) MV4; 11 0.082 12.6 0.2 RS4; 110.005 4.4 0.3 REH 0.006 4.2 1.1 Kasumi 0.016 10 1.2 HEL 0.034 11.3 >3.392.1.7 K562 0.046 20 >3.3 Jurkat 0.046 >10 >3.3

The data in Table 2 provides further evidence of per se activity byCompound 1 against all cell lines tested and particularly againstnon-MLL based AML, CML, erythroleukemia and ALL. The data also evidencesimproved efficacy of Compound 1 over SB216763 and GSK3-IX against allcell lines tested.

In Vivo Efficacy Experiments

Cultured cells (ATCC) are implanted subcutaneously in the rear flank offemale CD-1 nu/nu strain mice which have been acclimated for one week inthe animal facility after receipt from the vendor. Mice are randomizedinto groups of 10 mice per group and treatment begun when the mean tumorvolume reaches ˜100 mm³ Compound 1 is dosed IV. The tumors are measured2 times per week by electronic calipers to plot growth curves. Animalsare also monitored for fluctuations in body weight and survival.

Three cycles of 5 mg/kg of Compound 1 (injected IV) are given toanimals, each cycle separated by 7 days. Animals also receive 6 cyclesof Compound 1 (injected IV) given at 0.1 mg/kg and 1 mg/kg, each cycleseparated by 3.5 days. 30 mg/kg of the antimetabolite Arabinosylcytosine(injected IP) is given to animals every day for 14 consecutive days as acomparator control. p-value for each treatment group is determined bycomparison with the Captisol vehicle control group.

TABLE 3 Antitumor efficacy of Compound 1 in MV4; 11 leukemia xenograftsTumor Volume at day 33 Treatment Group Mean ± Standard Error (mm³)p-Value Captisol vehicle control 233 ± 23.2 — Compound 1 (5 mg/kg) 167 ±11.7 <0.01 administered once a week Compound 1 (0.1 mg/kg) 189 ± 22.3 —administered twice a week Compound 1 (1 mg/kg) 154 ± 15   <0.01administered twice a week Arabinosylcytosine (30 mg/kg) 129 ± 11.2 <0.001 administered every day for 14 days

The data in Table 3 evidences that the Compound 1 in vitro datademonstrating per se activity and improved efficacy, in the present testin comparison to the antimetabolite Arabinosylcytosine (injected IP), isalso seen in vivo.

Synthesis of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinyl-carbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineand pharmaceutically acceptable salts and solvates thereof areessentially as described in WO 2009/006043. As described below,synthesis is by common organic chemistry techniques known to one skilledin the art.

Preparation 1 2-imidazo[1,2-a]pyridin-3-yl-acetamide4,4-Dimethoxy-but-2-enoic acid ethyl ester

Add potassium carbonate (16.5 g, 120 mmol) to a solution of dimethoxyacetaldehyde (60% wt. in water) (15 mL, 100 mmol) and triethylphosphonoacetate (20 mL, 100 mmol) in 210 mL tetrahydrofuran and 30 mLwater. Stir the mixture at room temperature for 4 hours. Pour thereaction mixture into diethyl ether (200 mL) and wash with saturatedaqueous sodium chloride. Dry the organic phase over sodium sulfate andconcentrate under reduced pressure to provide the desired compound as ayellow oil (15.8 g, 90%).

¹H-NMR (300 MHz, CDCl₃): δ 6.77 (dd, J=15.9, 4.0 Hz, 1H), 6.13 (dd,J=15.9, 1.4 Hz, 1H), 4.95 (dd, J=4.0, 1.4 Hz, 1H), 4.22 (q, J=7.1 Hz,2H), 3.34 (s, 6H), 1.30 (t, J=7.1 Hz, 3H).

Imidazo[1,2-α]pyridin-3-yl-acetic acid ethyl ester

Heat a mixture of 4,4-dimethoxy-but-2-enoic acid ethyl ester (43.5 g,250 mmol) and p-toluenesulfonic acid (4.75 g, 25 mmol) in acetonitrile(240 mL) and water (15 mL) at reflux for 2 hours. Cool the reactionmixture to room temperature and add 2-aminopyridine (18.8 g, 200 mmol).Heat the mixture at reflux for 16 hours then cool to room temperature.Dilute the reaction mixture with ethyl acetate (1200 mL) and washsequentially with saturated aqueous sodium bicarbonate (600 mL×3) andsaturated aqueous sodium chloride (600 mL×2). Dry the organic phase oversodium sulfate and concentrate under reduced pressure to provide thedesired compound as a brown oil (30 g, 73%).

¹H-NMR (300 MHz, CDCl₃): δ 8.06 (d, J=6.6 Hz, 1H), 7.63 (d, J=9.1 Hz,1H), 7.56 (s, 1H), 7.20 (dd, J=8.9, 6.8 Hz, 1H), 6.84 (t, J=6.7 Hz, 1H),4.17 (q, J=7.3 Hz, 2H), 3.93 (s, 2H), 1.25 (t, J=7.3 Hz, 3H).

Amide Formation

Heat a solution of imidazo[1,2-α]pyridin-3-yl-acetic acid ethyl ester(30 g, 147 mmol) in NH₃/MeOH (7 N solution, 250 mL) at 85° C. in asealed tube for 15 hours. Cool the reaction mixture to room temperatureand concentrate under reduced pressure. Treat the residue withdichloromethane, sonicate, and filter the resulting precipitate toprovide the desired compound as a yellow solid (8.9 g, 35%).

¹H-NMR (300 MHz, DMSO): δ 8.30 (d, J=6.9 Hz, 1H), 7.62 (br s, 1H), 7.54(d, J=9.0 Hz, 1H), 7.42 (s, 1H), 7.21 (dd, J=7.7, 6.7 Hz, 1H), 7.18 (brs, 1H), 6.91 (t, J=6.8 Hz, 1H), 3.81 (s, 2H).

Preparation 29-Fluoro-7-methoxyoxalyl-3,4-dihydro-1H-[1,4]diazepino[6,7,1-hi]indole-2-carboxylicacid tert-butyl ester 2-Dibutoxymethyl-4-fluoro-1-nitro-benzene

Heat a solution of 5-fluoro-2-nitro-benzaldehyde (10 g, 59.17 mmol),butanol (20 mL, 219 mmol) and p-toluenesulfonic acid (600 mg, 3.15 mmol)in toluene (200 mL) at reflux for 2 hours in a flask equipped with aDean-Stark trap. Cool the reaction mixture to room temperature, dilutewith ethyl acetate (400 mL), and wash sequentially with saturatedaqueous sodium bicarbonate (300 mL×3) and saturated aqueous sodiumchloride (300 mL×2). Dry the organic phase over sodium sulfate andconcentrate under reduced pressure to provide the desired compound as apale yellow oil (17 g, 96%).

¹H-NMR (300 MHz, CDCl₃): δ 7.91 (dd, J=8.9, 4.9 Hz, 1H), 7.53 (dd,J=9.3, 2.9 Hz, 1H), 7.15-7.09 (m, 1H), 6.04 (s, 1H), 3.67-3.50 (m, 4H),1.63-1.54 (m, 4H), 1.44-1.32 (m, 4H), 0.92 (t, J=7.3 Hz, 6H).

5-Fluoro-1H-indole-7-carbaldehyde

Add vinylmagnesium bromide (1 M in tetrahydrofuran, 85.2 mL, 85.2 mmol)dropwise to a solution of 2-dibutoxymethyl-4-fluoro-1-nitro-benzene (8.5g, 28.4 mmol) in tetrahydrofuran (250 mL) at −78° C. Warm the reactionmixture −45° C. to −50° C. for 30 minutes, cool to −78° C., and addvinylmagnesium bromide (1 M in tetrahydrofuran, 85.2 mL, 85.2 mmol) dropwise. Warm the reaction mixture to −45° C. to −50° C. for 20 minutes,then add saturated aqueous ammonium chloride (300 mL). Warm the mixtureto room temperature and extract with diethyl ether (200 mL×2). Wash thecombined organic phases with saturated aqueous sodium chloride (400mL×2), dry over sodium sulfate, and concentrate under reduced pressure.Dissolve the residue in tetrahydrofuran (100 mL), add 0.5 N HCl (10 mL),and stir for 20 minutes. Dilute the mixture with diethyl ether (200 mL),wash sequentially with saturated aqueous sodium bicarbonate (200 mL×3)and saturated aqueous sodium chloride (200 mL×2). Dry the organic phaseover sodium sulfate and concentrate under reduced pressure. Subject theresidue to silica gel chromatography, eluting with 5% to 10% ethylacetate in hexanes to provide the desired compound as a pale yellowsolid (2.6 g, 56%).

¹H-NMR (300 MHz, CDCl₃): δ 10.07 (s, 1H), 10.05 (br s, 1H), 7.62 (d,J=7.6 Hz, 1H), 7.42-7.39 (m, 2H), 6.60 (d, J=5.4 Hz, 1H).

2-[(5-Fluoro-1H-indol-7-ylmethyl)-amino]-ethanol

Add 2-aminoethanol (1.93 mL, 32.0 mmol) followed by acetic acid (2.01mL, 48.0 mmol) to a solution of 5-fluoro-1H-indole-7-carbaldehyde (2.6g, 16.0 mmol) in 1,2-dichloroethane (40 mL). Stir at room temperaturefor 15 minutes. Add sodium triacetoxyborohydride (4.07 g, 19.2 mmol)portion wise. Stir the reaction mixture at room temperature for 3 hours.Add saturated aqueous sodium bicarbonate (100 mL) slowly followed by 1 NNaOH to pH ˜9. Extract with ethyl acetate (100 mL×3). Wash the organicphase with saturated aqueous sodium chloride (200 mL×2), dry over sodiumsulfate, and concentrate under reduced pressure to provide the desiredcompound as a pale yellow solid (3.2 g, 96%).

¹H-NMR (300 MHz, CDCl₃): δ 9.71 (br s, 1H), 7.24 (d, J=2.7 Hz, 1H), 7.19(dd, J=9.5, 2.3 Hz, 1H), 6.79 (dd, J=9.8, 2.2 Hz, 1H), 6.49 (dd, J=3.1,2.2 Hz, 1H), 4.15 (s, 2H), 3.77 (t, J=5.2 Hz, 2H), 2.84 (t, J=5.2 Hz,2H).

(5-Fluoro-1H-indol-7-ylmethyl)-(2-hydroxy-ethyl)-carbamic acidtert-butyl ester

Add a solution of di-tert-butyl dicarbonate (3.63 g, 16.65 mmol) intetrahydrofuran (40 mL) drop wise to a solution of2-[(5-fluoro-1H-indol-7-ylmethyl)-amino]-ethanol (3.15 g, 15.14 mmol) intetrahydrofuran (60 mL) at 0° C. Stir the reaction mixture at roomtemperature for 2 hours. Add ethyl acetate (200 mL) and wash withsaturated aqueous sodium chloride. Dry the organic phase over sodiumsulfate and concentrate under reduced pressure to provide the desiredcompound as a pale yellow oil (4.9 g, >100%).

¹H-NMR (300 MHz, CDCl₃): δ 10.17 (br s, 1H), 7.27-7.23 (m, 2H), 6.81(dd, J=9.4, 2.4 Hz, 1H), 6.50 (dd, J=2.9, 2.2 Hz, 1H), 4.67 (s, 2H),3.72 (br s, 2H), 3.33 (t, J=5.3 Hz, 2H), 1.50 (s, 9H).

Methanesulfonic acid2-[tert-butoxycarbonyl-(5-fluoro-1H-indol-7-ylmethyl)-amino]-ethyl ester

Add triethylamine (4.64 mL, 33.3 mmol) followed by methanesulfonylchloride (1.29 mL, 16.65 mmol) to a solution of(5-fluoro-1H-indol-7-ylmethyl)-(2-hydroxyethyl)-carbamic acid tert-butylester (4.9 g, assume 15.14 mmol) in dichloromethane (70 mL) at 0° C.Stir the reaction mixture for 30 minutes at 0° C. Dilute with ethylacetate (200 mL), wash with sequentially with saturated aqueous sodiumbicarbonate (200 mL×3) and saturated aqueous sodium chloride (200 mL×2).Dry the organic phase over sodium sulfate and concentrate under reducedpressure to provide the desired compound as a yellow brown oil (5.9 g,>100%).

¹H-NMR (300 MHz, CDCl₃): δ 10.07 (br s, 1H), 7.28-7.2 (m, 2H), 6.83 (dd,J=9.3, 2.3 Hz, 1H), 6.50 (dd, J=2.9, 2.2 Hz, 1H), 4.67 (s, 2H), 4.17 (t,J=5.5 Hz, 2H), 3.51 (t, J=5.6 Hz, 2H), 2.79 (s, 3H), 1.51 (s, 9H).

9-Fluoro-3,4-dihydro-1H-[1,4]diazepino[6,7,1-hi]indole-2-carboxylic acidtert-butyl ester

Add sodium hydride (60%) (666 mg, 16.65 mmol) in one portion to asolution of methanesulfonic acid2-[tert-butoxycarbonyl-(5-fluoro-1H-indol-7-ylmethyl)-amino]-ethyl ester(5.9 g, assume 15.14 mmol) in dimethylformamide (40 mL) at 0° C. Stirthe reaction mixture at 0° C. for 10 minutes and then at roomtemperature for 30 minutes. Add water (200 mL) slowly. Filter and drythe resulting yellow precipitate to provide the desired compound (4.14g, 94%).

¹H-NMR (300 MHz, CDCl₃): δ 7.15 (d, J=9.1 Hz, 1H), 7.07 (s, 1H), 6.78(dd, J=14.7, 8.8 Hz, 1H), 6.49 (d, J=3.1 Hz, 1H), 4.81 (s, 1H), 4.76 (s,1H), 4.25-4.23 (m, 2H), 3.94-3.83 (m, 2H), 1.49 (s, 9H).

9-Fluoro-7-methoxyoxalyl-3,4-dihydro-1H-[1,4]diazepino[6,7,1-hi]indole-2-carboxylicacid tert-butyl ester

Add oxalyl chloride (1.62 mL, 18.56 mmol) to a solution of9-fluoro-3,4-dihydro-1H-[1,4]diazepino[6,7,1-hi]indole-2-carboxylic acidtert-butyl ester (4.14 g, 14.28 mmol) in methyl tert-butyl ether (100mL) at −5° C. Warm the reaction mixture to room temperature over 1.5hours and then cool to −5° C. Add methanol (11.6 mL, 286 mmol) and stirat −5° C. for 30 minutes. Add saturated aqueous sodium bicarbonate (100mL) and extract with ethyl acetate (100 mL×3). Wash the combined organicphase sequentially with saturated aqueous sodium bicarbonate (200 mL×3)and saturated aqueous sodium chloride (200 mL×2). Dry the organic phaseover sodium sulfate and then concentrate under reduced pressure toprovide the title compound as a yellow solid (5.13 g, 93%).

¹H-NMR (300 MHz, CDCl₃): δ 8.38 (s, 1H), 8.04 (d, J=6.8 Hz, 1H), 6.89(dd, J=19.7, 8.6 Hz, 1H), 4.90 (s, 1H), 4.81 (s, 1H), 4.45-4.43 (m, 2H),4.05-3.93 (m, 2H), 3.95 (s, 3H), 1.42 (s, 9H).

Preparation 33-(9-Fluoro-1,2,3,4-tetrahydro-[1,4]diazepino[6,7,1-hi]indol-7-yl)-4-imidazo[1,2-a]-pyridin-3-yl-pyrrole-2,5-dionedihydrochloride

Add potassium tert-butoxide (4.58 g, 40.92 mmol) in one portion to asolution of9-fluoro-7-methoxyoxalyl-3,4-dihydro-1H-[1,4]diazepino[6,7,1-hi]indole-2-carboxylicacid tert-butyl ester (5.13 g, 13.64 mmol) and2-imidazo[1,2-a]pyridin-3-yl-acetamide (2.39 g, 13.64 mmol) indimethylformamide (80 mL). Stir the reaction mixture at room temperaturefor three hours. Add saturated aqueous ammonium chloride (200 mL) andextract with ethyl acetate (200 mL×3). Wash the combined organic phaseswith saturated aqueous sodium chloride (200 mL×3), dry over sodiumsulfate, and concentrate under reduced pressure. Dissolve the residue indichloromethane (20 mL) and add 4N HCl in dioxane (40 mL) drop wise,then stir at room temperature for 4 hours. Filter the resultingprecipitate and wash with diethyl ether to provide the title compound asa red solid (4.4 g, 68%).

MS (APCI): m/z=402 [C₂₂H₁₆FN₅O₂+H]⁺.

Example 17-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinyl-carbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepine

Add piperidine-1-carbonyl chloride (0.5 mL, 4.0 mmol) to a solution of3-(9-fluoro-1,2,3,4-tetrahydro-[1,4]diazepino[6,7,1-hi]indol-7-yl)-4-imidazo[1,2-a]pyridin-3-yl-pyrrole-2,5-dione(1.42 g, 3.0 mmol) and triethylamine (2.09 mL, 15.0 mmol) in methanol(80 mL). Stir at room temperature over night. Add triethylamine (1.04mL, 7.5 mmol) and piperidine-1-carbonyl chloride (0.5 mL, 4.0 mmol).Stir at room temperature for 5 hours. Add ethyl acetate (500 mL) andwash sequentially with saturated aqueous sodium bicarbonate (300 mL×3)and saturated aqueous sodium chloride (200 mL). Dry the organic phaseover sodium sulfate and concentrate under reduced pressure. Subject theresidue to silica gel chromatography, eluting with 0% to 3% methanol inethyl acetate to provide the title compound as a red solid (700 mg,45%).

m.p.=188-190° C.

MS (APCI): m/z=513 [C₂₈H₂₅FN₆O₃+H]⁺.

Example 27-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinyl-carbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepinemethanesulfonate

Heat a slurry of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinyl-carbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepine(500 mg, 0.976 mmol) in methanol (2.5 mL) to 64° C. Add a solution ofmethanesulfonic acid (64 μL, 0.976 mmol) in methanol (1.0 mL) over 5minutes. Stir the mixture at 64° C. for 15 minutes and then addisopropanol (5.0 mL) over 30 minutes. Allow the resulting slurry to coolto room temperature over 1 hour and then stir at room temperature for 4hours. Filter the slurry, wash with isopropanol, and dry under reducedpressure at 42° C. to provide the title compound as an orange solid (478mg, 88.5% (adjusted for 9.9% volatiles in starting material and 1.0%volatiles in product)).

m.p.=282.3° C. (DSC)

Example 37-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinyl-carbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineethanolate

Heat a slurry of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinyl-carbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepine(2.0 g, 3.9 mmol) in ethanol (30 mL) to 70° C. Add 5M HCl (0.73 mL) allat once. Stir the mixture at 70° C. for 10 minutes and then add 1N NaOH(3.63 mL) over 3 minutes. Stir the mixture at 70° C. for 2 hours. Allowthe resulting slurry to cool to room temperature over 1 hour and thenstir at room temperature for 3.5 hours. Filter the slurry, wash withethanol, and dry under reduced pressure at 42° C. to provide the titlecompound as an orange solid (1.84 g, 92% (adjusted for 7.5% volatiles instarting material and 7.7% volatiles in product)).

m.p.=179.4° C. (DSC)

Powder X-ray Principal Peaks (Degrees 2 Theta, Intensity): 8.989°, 100%;9.787°, 48.7%; 12.846°, 20.0%; and 7.444°, 17.5%.

Example 47-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinyl-carbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepinehydrate I

Heat a slurry of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinyl-carbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineethanolate (198.5 mg) in water (10 mL) to 80° C. for 2.75 hours. Add3.11 mL of 1N HCl. When the temperature has returned to 80° C., rapidlyadd 3.11 mL of 1N NaOH. Allow the temperature to remain at 80° C. forapproximately 15 minutes then allow the suspension to cool to roomtemperature. Collect the solid using vacuum filtration through Whatman#1 paper and allow to dry loosely covered over night. Powder X-rayPrincipal Peaks (Degrees 2 Theta, Intensity): 12.089°, 100%; 10.485°,83.6%; 13.227°, 56.0%; and 7.660°, 8.0%.

Example 57-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinyl-carbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepinehydrate II

Heat a slurry of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinyl-carbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineethanolate (200.6 mg) in water (25 mL) to 75° C. for 0.5 hours. Add 0.72mL of 1N HCl and continue to heat for 0.75 hours. Rapidly add 0.72 mL of1N NaOH. Allow the suspension to cool to room temperature. Collect thesolid using vacuum filtration through Whatman #1 paper, rinse with 20 mLdeionized water and allow to dry loosely covered for 2 days.

Powder X-ray Principal Peaks (Degrees 2 Theta, Intensity): 6.878°, 100%;5.732°, 58.7%; 11.550°, 82.8%; 18.426°, 20.7%; and 10.856°, 44.2%.

Example 67-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinyl-carbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepinedihydrate

Heat a slurry of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)-9-fluoro-1,2,3,4-tetrahydro-2-(1-piperidinyl-carbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineethanolate (200.8 mg) in water (25 mL) to 75° C. for 0.67 hours. Add0.72 mL of 1N HCl and continue to heat for 1.75 hours. Add 0.1N NaOH in1 mL increments every 5 minutes until 7.2 mL have been added. After thelast addition, allow the suspension to remain at 75° C. for 0.67 hoursand then allow the suspension to cool to room temperature. Collect thesolid using vacuum filtration through Whatman #1 paper, rinse with 20 mLdeionized water and allow to dry loosely covered for 2 days.

Powder X-ray Principal Peaks (Degrees 2 Theta, Intensity): 5.498°, 100%;22.149°, 100%; 14.921°, 32.9%; 11.399°, 36.7%; and 11.019°, 20.5%.

Compound 1 is preferably formulated as a pharmaceutical compositionprior to administration to a patient. Useful formulations compriseCompound 1 or a pharmaceutically acceptable salt or solvate thereof andSBE7-β-CD. The compound SBE7-β-CD is a sulfobutyl ether ofβ-cyclodextrin described in U.S. Pat. No. 5,134,127. It is sold underthe trade name CAPTISOL®. Particular formulations are described in thefollowing Formulation Examples.

A useful pharmaceutical composition may be prepared by dissolvingCompound 1 or a pharmaceutically acceptable salt or solvate thereof (50mg/mL) in 2-pyrrolidone (SOLUPHOR®-P). This solution is then dilutedwith an aqueous solution of SBE7-β-CD (30% by volume) and poloxamer 188(Lutrol®-F 68) (10% by volume).

Formulation Example 1

Prepare a first solution by adding 30.0 g SBE7-β-CD to 71.25 mL of waterand stir or agitate until completely dissolved. Add 10.0 g poloxamer 188and continue stirring until completely dissolved. Prepare a secondsolution by adding Compound 1 ethanolate to 2-pyrrolidone according tothe following formula: mL 2-pyrrolidone=(actual Compound 1 ethanolate wt(mg)/50 mg/mL)×0.5. Add the first solution to the second solution.Filter the resulting solution through a 0.2 μm SUPOR® (hydrophilicpolyethersulfone) filter (Pall Corporation) into a dust free container.

A further pharmaceutical composition embodiment is prepared by combiningCompound 1 or a pharmaceutically acceptable salt or solvate thereof inan equimolar amount of a pharmaceutically acceptable acid in water. Thismixture is then combined with at least one molar equivalent of SBE7-β-CDas an aqueous solution. Preferred pharmaceutically acceptable acidsinclude HCl, HBr, sulfuric acid and methanesulfonic acid. The use of HClis especially preferred.

Formulation Example 2

Prepare a first solution by adding 20.0 g SBE7-β-CD to 80.0 mL of waterand stir or agitate until completely dissolved. Add this solution toCompound 1 ethanolate according to the following formula: mL of firstsolution=(actual Compound 1 ethanolate wt (mg)/20 mg/mL)-(actualCompound 1 ethanolate wt (mg)/1200 mg/mL)-(actual Compound 1 ethanolatewt (mg)×0.00195107 mL of 1N HCl/mg Compound 1 ethanolate). Add 1N HClaccording to the following calculation: mL of 1N HCl to add=(actualCompound 1 ethanolate wt (mg)×0.00195107 mL of 1N HCl/mg Compound 1ethanolate). Stir or bath sonicate until all compound has dissolved.

A preferred pharmaceutical composition embodiment is prepared by adding1 molar equivalent of Compound 1 or a pharmaceutically acceptable saltor a solvate thereof to an aqueous solution of at least 1 molarequivalent of SBE7-β-CD at a pH below 5.5 (initial solution pH),optionally in the presence of a pharmaceutically acceptable buffer, andmixing until the Compound 1 or a pharmaceutically acceptable salt orsolvate thereof has dissolved. The pH is then adjusted to between 2.5and 3.5 with a pharmaceutically acceptable base (final solution pH).This resulting solution formulation may be administered to a patientdirectly, or the solution may preferably be lyophilized to provide asolid formulation capable of reconstitution with water.

The SBE7-β-CD may be present in the range of 1 molar equivalent up to anamount required to administer no more than 13.4 gm of SBE7-β-CD to apatient in a day. A preferred amount of SBE7-β-CD is from 1.0 to 4.0molar equivalents, more preferred is from 2.0 to 3.0 molar equivalents,and from 2.5 to 2.7 molar equivalents relative to Compound 1 isespecially preferred.

Although any initial solution pH below 5.5 is acceptable, an initialsolution pH below 3.0 is preferred, an initial solution pH in the rangeof 1.0 to 2.0 is more preferred, and an initial solution pH of between1.2 and 1.4 is most preferred. The target initial solution pH isachieved by the addition of any pharmaceutically acid capable ofadjusting the pH of the solution to a pH less than 5.5. The use ofhydrochloric acid is preferred.

The formulation may optionally contain a pharmaceutically acceptablebuffer. Pharmaceutically acceptable buffers are those compounds employedby one skilled in the pharmaceutical formulation arts to stabilize thepH of a final solution in a particular pH range. Pharmaceuticallyacceptable buffers include phosphate buffers as well as citric acid,glycine, and tartaric acid or pharmaceutically acceptable salts thereof.Pharmaceutically acceptable salts of these acids include the sodium andpotassium salts. It is preferred that a pharmaceutically acceptablebuffer is present in the formulation. Tartaric acid is a preferredpharmaceutically acceptable buffer.

It is important that the Compound 1 dissolve completely before the pH isadjusted to the final solution pH. Dissolution may be assisted by anymechanical mixing means or by adjusting the temperature of the solutionif necessary or desired. Stirring the solution at room temperature ispreferred.

The final solution pH is achieved by the addition of anypharmaceutically acceptable base capable of adjusting the pH of thesolution to a pH in the range of 2.5 to 3.5. The use of sodium hydroxideis preferred. The final solution pH may be in the range of 2.5 to 3.5,but is preferably in the range of 2.5 to 3.1. A final solution pH in therange of 2.7 to 3.1 is most preferred. Once the final solution pH hasbeen achieved, the solution may be lyophilized if necessary or desiredunder standard lyophilization conditions to provide a solidpharmaceutical composition suitable for reconstitution with water.

Formulation Example 3

Prepare a solution of 0.15 g tartaric acid and 12 g (5.55 mmol)SBE7-β-CD in 70 mL of water. Add 5 mL of 1.0 N HCl and mix at roomtemperature. Add 1.1 g (2.15 mmol) Compound 1 ethanolate and stir atroom temperature until dissolved. Add 1N sodium hydroxide to a pH ofabout 2.9. Add sufficient water to achieve a final volume of 100 mL.Lyophilize this solution to provide an amorphous orange-red solid.

I claim:
 1. A method of treating a patient suffering from translocatedmixed lineage leukemia based acute myelogenous leukemia comprisingadministering to a patient in need of treatment an effective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)9-flouro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor pharmaceutically acceptable salt or solvate thereof.
 2. A method oftreating a patient suffering from translocated mixed lineage leukemiabased acute lymphoid leukemia comprising administering to a patient inneed of treatment an effective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)9-flouro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor pharmaceutically acceptable salt or solvate thereof.
 3. A method oftreating a patient suffering from a non-mixed lineage leukemia basedchronic myeloproliferative disorder which is erythroleukemia comprisingadministering to a patient in need of treatment an effective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)9-flouro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor pharmaceutically acceptable salt or solvate thereof.
 4. A method oftreating a patient suffering from a non-mixed lineage leukemia basedchronic myeloproliferative disorder which is chronic myelogenousleukemia comprising administering to a patient in need of treatment aneffective amount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)9-flouro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor pharmaceutically acceptable salt or solvate thereof.
 5. A method oftreating a patient suffering from a non-mixed lineage leukemia basedchronic myeloproliferative disorder which is acute lymphoid leukemiacomprising administering to a patient in need of treatment an effectiveamount of7-(2,5-dihydro-4-imidazo[1,2-a]pyridine-3-yl-2,5-dioxo-1H-pyrrol-3-yl)9-flouro-1,2,3,4-tetrahydro-2-(1-piperidinylcarbonyl)-pyrrolo[3,2,1-jk][1,4]benzodiazepineor pharmaceutically acceptable salt or solvate thereof.